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C. Sano, A. Namavari, M. Bakir, S. Jain; Neurotrophin and Nerve Guidance Protein Presence and Expression in Preserved Donor Cornea. Invest. Ophthalmol. Vis. Sci. 2010;51(13):6214.
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Abnormalities in the extent and distribution of corneal nerves in the graft persist for several years after corneal transplant. In these experiments our goal is to determine the changes in the expression of neurotrophins and nerve guidance proteins in optisol GS preserved corneal donor tissue.
2.0-mm diameter murine corneal buttons were trephined and immediately placed in sterile optisol GS preservative solution. Corneas (n=16/timepoint) were analyzed at time 0 (immediately after trephination), day 3 and day 7. Neurotrophins (NGF, GDNF, BDNF, CNTF, NT-3, NT4/5), Nerve guidance proteins (EphB1, ephrinb1), vimentin and alpha-smooth muscle actin levels were assayed by quantitative RT-PCR (8 corneas/timepoint) and quantitative Western analyses (LiCor).
EphB1/ephrinB1 were differentially expressed. EphB1 was upregulated in corneal buttons by 3% on day 3, however was downregulated by 18% on day 7. EphrinB1 was downregulated by 25% between day 0 and 3, however was upregulated by 7% by day 7 compared to day 3. Of the neurotrophins, GDNF and NGF showed upregulation on day 3 followed by downregulation on day 7. Western analysis showed NGF levels increasing levels from 0-7 days (94% increase on day 3 and a 97% increase by day 7).
Expression of neurotrophins and nerve guidance proteins changes with the duration of corneal graft preservation.
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